Tumour necrosis factor (TNF) was first identified as a factor found in the serum of Bacillus Calmette-Guerin treated mice which caused haemorrhagic regression of certain transplanted tumours and had cytolytic activity on several transformed cell lines in vitro (Carswell et al, PNAS 72, 3666-3670; Helson et al, 1975, Nature 258, 731-732). TNF, a product of activated macrophages, has subsequently been shown to be a primary mediator in the pathology of endotoxic shock (Tracey et al 1986, Science 234, 470-474). In addition to its pathological effects TNF also has a central role in host defences against viral, bacterial and parasitic pathogens.
The cellular targets of TNF important in host defence include neutrophils, eosinophils, monocyte/macrophages and lymphocytes. Within this context TNF is a major mediator of neutrophil activation. TNF stimulates enhanced phagocytosis (Shalaby et al 1985, J. Immunol., 135, 2069-2073), enhanced production of superoxide anions (Teujiimoto et al, 1986, Biochem. Biophys. Res. Commun., 137, 1094-1100), release of lysozyme and hydrogen peroxide and causes neutrophil degranulation (Klebanoff et al, 1986, J. Immunol., 136, 4220-4225). Neutrophils also show enhanced microbiocidal and tumouricidal activity when stimulated by TNF (Shalaby et al, 1985, J. Immunol., 135, 2069-2073; Djeu et al, 1986, J. Immunol., 137, 2980-2984; Blanchard et al, 1989, J. Leuk. Biol., 45, 538-545). It has been hypothesized that the cytostatic effect of TNF is mediated by high local concentrations of hydrogen peroxide produced by neutrophils (Shau 1986, J. Immunol., 141, 234-240).
TNF pretreatment enhances the response of neutrophils to N-formyl-L-methionyl-L-leucyl-L-phenylalanine (F-met-leu-phe) and phorbol myristate acetate through specific receptors (Ferrante et al 1988, Int. Arch. Allergy AppI. Immunol., 86, 82-91). Neutrophils accumulate at sites of inflammation, caused in part by the increased expression of complement receptors by TNF (Berger et al 1988, Blood 71, 151-158). Further TNF causes neutrophil emigration into skin (Cybulsky et al 1988, J. Immunol. 140, 3144-3149).
Neutrophil function is known to be depressed in a number of viral, bacterial and parasitic infections (Abramson and Mills, 1988, Rev. Infect. Dis., 10, 326-341; Ferrante et al, 1989, Immunol. Letts., 22, 301-6). Depressed neutrophil function has, for example, been described in Acquired Immune Deficiency Syndrome (Thorsen et al, 1989, AIDS, 3, 651-653; Ellis et al, 1988, J. Infect. Dis., 158, 1268-1276; Murphy et al, 1988, J. Infect. Dis., 158, 627-630). Clearly TNF, which appears to play an important role in neutrophil activation both in vitro and in vivo as described above, given exogenously has the potential to overcome these neutrophil defects. The administration of TNF or indeed overproduction of TNF is, however, associated with severe side effects and the manifestation of pathology such as thrombocytopaenia, lymphocytopaenia, hepatotoxicity, renal impairment and hypertension.
The present inventors have identified novel peptides derived from the primary amino acid sequence of human TNF which stimulate neutrophil activity. These peptides have indicated that the region of amino acids 54 to 94 of human TNF has previously undiscovered neutrophil stimulating activity. This observation has important clinical applications as treatment with such peptides would be expected to restore depressed or aberrant neutrophil activity, but would not be expected to cause the severe side effects associated with-the therapeutic use of the whole TNF molecule.
The present invention provides a method of detecting a site of inflammation in a subject which method comprises administering to the subject a detectably labeled peptide, the peptide comprising a sequence selected from the group consisting of LFKGQGCPSTHVLLTHTISRI (SEQ ID NO. 6), GLYLIYSQVLFKGQG (SEQ ID NO. 10), HVLLTHTISRIAVSYQTKVNLL (SEQ ID NO. 11), PSTHVLLTHTI (SEQ ID NO. 15), PSAHVLLTHTI (SEQ ID NO. 17), and PSTHVLITHTI (SEQ ID NO. 18); and detecting the labeled peptide.
The present invention also provides a method of diagnosing a disease state selected from the group consisting of a bacterial infection, a fungal infection, a parasitic infection, a viral infection, Group B streptococcal disease, osteomyelitis, pneumonia, bronchitis, chronic obstructive pulmonary disease, inflammatory bowel disease, atypical appendicitis, Acquired Immune Deficiency Syndrome, a granulomatous disease, respiratory distress syndrome, cystic fibrosis and rheumatoid arthritis, which method comprises administering to a subject a detectably labeled peptide, the peptide comprising a sequence selected from the group consisting of LFKGQGCPSTHVLLTHTISRI (SEQ ID NO. 6), GLYLIYSQVLFKGQG (SEQ ID NO. 10), HVLLTHTISRIAVSYQTKVNLL (SEQ ID NO. 11), PSTHVLLTHTI (SEQ ID NO. 15), PSAHVLLTHTI (SEQ ID NO. 17), and PSTHVLITHTI (SEQ ID NO. 18); and detecting the labeled peptide.
The present invention relates to a peptide which primes neutrophils for superoxide production and an enhanced respiratory burst following treatment with N-formyl-L-methionyl-L-leucyl-L-phenylalanine, wherein the peptide is of the general formula:
X1-X2-X3-X4-X5-X6-X7 in which
X1 is null, Cys or R1 
X2 is null, Cys, R1, or A1-A2-A3-A4-A5-A6-A7
xe2x80x83in which A1 is Leu, Ile, Val or Met
A2 is Phe, Tyr, Trp or His
A3 is Lys, Arg or His
A4 is Gly or Ala
A5 is Gln or Asn
A6 is Gly or Ala
A7 is Cys
X3 is Cys, R1 or A8-A9-A10
xe2x80x83in which A8 is Pro or Nxcex1-alkylamino acid
A9 is Ser or Thr
A10 is Thr or Ser or Ala or Gly
X4 is A11-A12-A13-A14-A15-A16-A17-A18
xe2x80x83in which A11 is His, Lys or Arg
A12 is Val, Ile, Leu or Met
A13 is Leu, Ile Val or Met
A14 is Ile, Leu, Val or Met
A15 is Thr or Ser
A16 is His, Lys or Arg
A17 is Thr or Ser
A18 is 16, Leu, Val or Met
X5 is Cys, R2 or A19-A20-A21
xe2x80x83in which A19 is Ser or Thr
A20 is Arg, Lys or His
A21 is Ile, Leu, Val or Met
X6 is null, Cys, R2 or A22-A23-A24-A25-A26-A27-A28-A29-A30-A31-A32
xe2x80x83in which A22 is Ala or Gly
A23 is Val, Ile, Leu or Met
A24 is Ser or Thr
A25 is Tyr, Phe, Trp or His
A26 is Glu or Asp
A27 is Thr or Ser
A28 is Lys, Arg or His
A29 is Val, Ile, Leu or Met
A30 is Asn or Gln
A31 is Leu, Ile, Val or Met
A32 is Leu, Ile, Val or Met
X7 is null, Cys or R2
R1 is H or Rxe2x80x94CO, where R is H, straight, branched or cyclic alkyl up to C20, optionally containing double bonds and/or substituted with halogen, nitro, amino, hydroxy, sulfo, phospho or carboxyl groups which may be substituted themselves or aralkyl or aryl optionally substituted as listed for the alkyl or R1 is glycosyl, nucleosyl or lipoyl and R1 is absent when the amino acid adjacent is an unsubstituted desamino-derivative; R2 is xe2x80x94NR12R13, wherein R12 and R13 are independently H, straight, branched or cyclic alkyl, aralkyl or aryl optionally substituted as defined for R1 or R2 is N-glycosyl or N-lipoyl, or R2 is xe2x80x94OR14, where R14 is H straight, branched or cyclic alkyl, aralkyl or aryl, optionally substituted as defined for R1 or R2 is xe2x80x94O-glycosyl, or xe2x80x94O-lipoyl or R2 is absent when the adjacent amino acid is a dicarboxy derivative of cysteine or a homologue thereof or the peptide is in a Nxe2x80x94C cyclic form, with the proviso that:
X1 is always and only null when X2 is R1, Cys or null
X2 is always and only null when X3 is R1 or Cys
X6 is always and only null when X5 is R2 or Cys
X7 is always and only null when X6 is R2 or Cys or null.
In a preferred embodiment of the present invention X1 is null, X2 is R1, X3 is A8-A9-A10, X5 is R2 and X6 and X7 are null. It is further preferred that A8 is Pro, A9 is Ser, A10 is Thr, A11 is His, A12 is Val, A13 is Leu, A14 is Leu, A15 is Thr, A16 is His, A17 is Thr and A18 is Ile, or A8 is Pro, A9 is Ser, A10 is Thr, A11 is His, A12 is Val, A13 is Leu, A14 is Ile, A15 is Thr, A16 is His, A17 is Thr and A18 is Ile, or A8 is Pro, A9 is Ser, A10 is Ala, A11 is His, A12 is Val, A13 is Leu, A14 is Leu, A15 is Thr, A16 is His, A17 is Thr and A18 is Ile.
In another preferred embodiment of the present invention X1 is R1, X2 is A1-A2-A3-A4-A5-A6-A7, X3 is A8-A9-A10, X5 is A19-A20-A21, X6 is R2 and X7 is null. It is further preferred that A1 is Leu, A2 is Phe, A3 is Lys, A4 is Gly, A5 is Gln, A6 is Gly, A7 is Cys, A8 is Pro, A9 is Ser, A10 is Thr, A11 is His A12 is Val, A13 is Leu, A14 is Leu, A15 is Thr, A16 is His, A17 is Thr, A18 is Ile, A19 is Ser, A20 is Arg and A21 is Ile.
In yet another preferred embodiment of the present invention X1 and X2 are null, X3 is R1, X5 is A19-A20-A21, X6 is A22-A23-A24-A25-A26-A27-A28-A29-A30-A31-A32 and X7 is R2. It is further preferred that A11 is His, A12 is Val, A13 is Leu, A14 is Leu, A15 is Thr, A16 is His A17 is Thr, A18 is Ile, A19 is Ser, A20 is Arg, A21 is Ile, A22 is Ala, A23 is Val, A24 is Val, A25 is Ser, A26 is Tyr, A27 is Glu, A28 is Lys, A29 is Val, A30 is Asa, A31 is Leu and A32 is Leu.
In a second aspect the present invention consists in a peptide which primes neutrophils for superoxide production and an enhanced respiratory burst following treatment with N-formyl-L-methionyl-L-leucyl-L-phenylalanine, wherein the peptide is of the general formula:
xe2x80x83B1-Y1-Y2-Y3-Y4-Y5-Y6-Y7-Y8-Y9-Y10-Y11-Y12-Y13-Y14-Y15-B2
in which Y1 is Gly or Ala
Y2 is Leu or Ile or Val or Met
Y3 is Tyr or Phe or Trp or His
Y4 is Leu or Ile or Val or Met
Y5 is Ile or Leu or Val or Met
Y6 is Tyr or Phe or Trp or His
Y7 is Ser or Thr
Y8 is Gln or Asn
Y9 is Val or Ile or Leu or Met
Y10 is Leu or Val or Ile or Met
Y11 is Phe or Tyr or Trp or His
Y12 is Lys or Arg or His
Y13 is Gly or Ala
Y14 is Asn or Gln
Y15 is Gly or Ala
B1 is H or R-CO, where R is H, straight, branched or cyclic alkyl up to C20, optionally containing double bonds and/or substituted with halogen, nitro, amino, hydroxy, sulfo, phospho or carboxyl groups (which may be substituted themselves or aralkyl or aryl optionally substituted as listed for the alkyl or B1 is glycosyl, nucleosyl or lipoyl and B1 is absent when the amino acid adjacent is an unsubstituted desamino-derivative; B2 is xe2x80x94NR12R13, wherein R12 and R13 are independently H, straight, branched or cyclic alkyl, aralkyl or aryl optionally substituted or defined for B1 or B2 is N-glycosyl or N-lipoyl, or B2 is xe2x80x94OR14, where R14 is H straight, branched or cyclic alkyl, aralkyl or aryl, optionally substituted as defined for B1 or B2 is xe2x80x94O-glycosyl, or xe2x80x94O-lipoyl or B2 is absent when the adjacent amino acid is a dicarboxy derivative of cysteine or a homologue thereof or the peptide is in a Nxe2x80x94C cyclic form.
As will be appreciated by those skilled in the art from the description which follows the present inventors have demonstrated that the region of human TNF from amino acid 54 to amino acid 94 plays an important functional role in the stimulation of neutrophils. Further, the present inventors have produced 6 peptides namely peptides 304, 308, 309, 395, 418 and 419 (as referred to herein) which have neutrophil stimulating activity.
Armed with this information and with the aid of co-ordinates of the crystalline structure of TNF at 2.6 A as disclosed by Eck and Sprang, 1989 (J. Biol. Chem., 264: 18795-17605), the person skilled in the art will be able to design non-peptide structures which, in 3 dimensional terms mimic the peptides of the present invention. It is believed that these non-peptide structures will also mimic the physiological effects of the peptides of the present invention. It is intended that such non-peptide structures are included within the scope of the present invention. Changes to the TNF molecule in these regions using eg. site directed mutagenesis would also be expected to affect neutrophil activation. A schematic representation of the three dimensional structure of TNFxcex1 is shown in FIG. 8.
Accordingly in a third aspect the present invention consists in a compound the three dimensional structure of which substantially corresponds to the three dimensional structure of the peptide of the first or second aspects of the present invention, the compound being characterized in that the compound is capable of eliciting superoxide production by neutrophils and of priming neutrophils for an enhanced respiratory burst following treatment with N-formyl-L-methionyl-L-leucyl-L-phenylalanine.
In a further aspect, the present invention consists in a method of treating a subject having depressed neutrophil function, the method comprising administering to the subject a therapeutic amount of the peptide of the first aspect of the present invention.
In a preferred embodiment of this aspect of the present invention the subject is suffering from acquired immune deficiency syndrome.
Peptide 308 (SEQ ID NO. 10), through selective effects on neutrophil degranulation may be administered to individuals suffering from inflammatory syndromes e.g. rheumatoid arthritis, adult respiratory distress syndrome.
It will be appreciated by those skilled in the art that a number of modifications may be made to the peptide of the present invention without deleteriously effecting the biological activity of the peptide. This may be achieved by various changes, such as insertions, deletions and substitutions (e.g., sulfation, phosphorylation, nitration, halogenation), either conservative or non-conservative (e.g., W-amino acids, desamino acids) in the peptide sequence where such changes do not substantially altering the overall biological activity of the peptide. By conservative substitutions the intended combinations are:
G, A; V, I, L, M; D, E; N, Q; S, T; K, R, H; F, Y, W. H; and P, N(-alkylamino acids.
It may also be possible to add various groups to the peptide of the present invention to confer advantages such as increased potency or extended half-life in vivo, without substantially altering the overall biological activity of the peptide.
The term peptide is to be understood to embrace peptide bond replacements and/or peptide mimetics, i.e. pseudopeptides, as recognised in the art (see for example: Proceedings of the 20th European Peptide Symposium, edt. G. Jung. E. Bayer, pp. 289-336, and references therein), as well as salts and pharmaceutical preparations and/or formulations which render the bioactive peptide(s) particularly suitable for oral, topical, nasal spray, ocular pulmonary, I.V., subcutaneous, as the case may be, delivery. Such salts, formulations, amino acid replacements and pseudopeptide structures may be necessary and desirable to enhance the stability, formulation, deliverability (e.g., slow release, prodrugs), or to improve the economy of production, and they are acceptable, provided they do not negatively affect the required biological activity of the peptide.
Apart from substitutions, three particular forms of peptide mimetic and/or analogue structures of particular relevance when designating bioactive peptides, which have to bind to a receptor while risking the degradation by proteinases and peptidases in the blood, tissues and elsewhere, may be mentioned specifically, illustrated by the following examples: Firstly, the inversion of backbone chiral centres leading to D-amino acid residue structures may, particularly at the N-terminus, lead to enhanced stability for proteolytical degradation while not impairing activity. An example is given in the paper xe2x80x9cTritriated D-ala1-Peptide T Bindingxe2x80x9d, Smith, C. S. et al, Drug Development Res. 15, pp. 371-379 (1988). Secondly, cyclic structure for stability, such as N to C interchain imides and lactames (Ede et al in Smith and Rivier (Eds) xe2x80x9cPeptides: Chemistry and Biologyxe2x80x9d, Escom, Leiden (1991), p268-270), and sometimes also receptor binding may be enhanced by forming cyclic analogues. An example of this is given in xe2x80x9cConfirmationally restricted thymopentin-like compoundsxe2x80x9d, U.S. Pat. No. 4,457,489 (1985), Goldstein, G. et al. Finally, the introduction of ketomethylene, methylsulfide or retroinverse bonds to replace peptide bonds, i.e. the interchange of the CO and NH moieties may both greatly enhance stability and potency. An example of the latter type is given in the paper xe2x80x9cBiologically active retroinverse analogues of thymopentinxe2x80x9d, Sisto A. et al in Rivier, J. E. and Marshall, G. R. (eds.) xe2x80x9cPeptides, Chemistry, Structure and Biologyxe2x80x9d, Escom, Leiden (1990), p.722-773.
The peptides of the invention can be synthesized by various methods which are known in principle, namely by chemical coupling methods (cf. Wunsch, E.: xe2x80x9cMethoden der organischen Chemiexe2x80x9d, Volume 15, Band 1+2, Synthese von Peptiden, Thieme Verlag, Stuttgart (1974), and Barrany, G.;
Merrifield, R. B: xe2x80x9cThe Peptidesxe2x80x9d, eds. E. Gross, J. Meienhofer., Volume 2, Chapter 1, pp. 1-284, Academic Press (1980)), or by enzymatic coupling methods (cf. Widmer, F., Johansen, J. T., Carlsberg Res. Commun., Volume 44, pp. 37-46 (1979), and Kullmann, W.: xe2x80x9cEnzymatic Peptide Synthesisxe2x80x9d, CRC Press Inc., Boca Raton, Fla. (1987), and Widmer, F., Johansen, J. T. in xe2x80x9cSynthetic Peptides in Biology and Medicine:, eds., Alitalo, K., Partanen, P., Vatieri, A., pp. 79-86, Elsevier, Amsterdam (1985)), or by a combination of chemical and enzymatic methods if this is advantageous for the process design and economy.
It will be seen that one of the alternatives embraced in the general formula set out above is for a cysteine residue to be positioned at both the amino and carboxy terminals of the peptide. This will enable the cyclisation of the peptide by the formation of di-sulphide bond.
It is intended that such modifications to the peptide of the present invention which do not result in a decrease in biological activity are within the scope of the present invention.
As would be recognized by those skilled in the art there are numerous examples to illustrate the ability of anti-idiotypic (anti-Ids) antibodies to an antigen to function like that antigen in its interaction with animal cells and components of cells. Thus, anti-Ids to a peptide hormone antigen can have hormone-like activity and interact specifically with the receptors to the hormone. Conversely, anti-Ids to a receptor can interact specifically with a mediator in the same way as the receptor does. (For a review of these properties see: Gaulton, G. N. and Greane, M. I. 1986. Idiotypic mimicry of biological receptors, Ann. Rev. Immunol. 4, 253-280; Sege, K and Peterson, P. A., 1978. Use of anti-idiotypic antibodies as cell surface receptor probes. Proc. Natl. Acad. Sci. U.S.A. 75, 2443-2447).
As might be expected from this functional similarity of anti-Id and antigen, anti-Ids bearing the internal image of an antigen can induce immunity to such an antigen. (This nexus is reviewed in Hiernaux, J. R. 1988. Idiotypic vaccines and infectious diseases. Infect. Immun. 56, 1407-1413.)
As will be appreciated by persons skilled in the art from the disclosure of this application it will be possible to produce anti-idiotypic antibodies to the peptide of the present invention which will have similar biological activity. It is intended that such anti-idiotypic antibodies are included within the scope of the present invention.
Accordingly, in a fourth aspect the present invention consists in an anti-idiotypic antibody to the peptide of the first aspect of the present invention, the anti-idiotypic antibody being capable of inducing macrophage and/or neutrophil activation.
The individual specificity of antibodies resides in the structures of the peptide loops making up the Complementary Determining Regions (CDRs) of the variable domains of the antibodies. Since in general, the amino acid sequences of the CDR peptide loops of an anti-Id are not identical to or even similar to the amino acid sequence of the peptide antigen from which it was originally derived, it follows that peptides whose amino acid sequence is quite dissimilar, in certain contexts can take up a very similar three-dimensional structure. The concept of this type of peptide, termed a xe2x80x9cfunctionally equivalent sequencexe2x80x9d or mimotope by Geyson is familiar to those expert in the field. (Geyson, H. M. et al 1987. Strategies for epitope analysis using peptide synthesis. J. Immun. Methods. 102, 259-274).
Moreover, the three-dimensional structure and function of the biologically active peptides can be simulated by other compounds, some not even peptidic in nature, but which mimic the activity of such peptides. This field of science is summarised in a review by Goodman, M. (1990). (Synthesis, spectroscopy and computer simulations in peptide research. Proc. 11th American Peptide Symposium published in Peptides-Chemistry, Structure and Biology pp 3-29. Ed Rivier, J. E. and Marshall, G. R. Publisher ESCOM.)
As will be recognized by those skilled in the art, armed with the disclosure of this application, it will be possible to produce peptide and non-peptide compounds having the same three-dimensional structure as the peptide of the present invention. These xe2x80x9cfunctionally equivalent structuresxe2x80x9d or xe2x80x9cpeptide mimicsxe2x80x9d will react with antibodies raised against the peptide of the present invention and may also be capable of stimulating macrophages and/or neutrophils. It is intended that such xe2x80x9cpeptide mimicsxe2x80x9d are included within the scope of the present invention.
Accordingly, in a fifth aspect the present invention consists in a compound the three-dimensional structure of which is similar as a pharmacophore to the three-dimensional structure of the peptide of the first aspect of the present invention, the compound being characterized in that it reacts with antibodies raised against the peptide of the first aspect of the present invention and that the compound is capable of activating macrophages and/or neutrophils.
More detail regarding pharmacophores can be found in Bolin et al. p 150, Polinsky et al. p 287, and Smith et al. p 485 in Smith and Rivier (Eds) xe2x80x9cPeptides: Chemistry and Biologyxe2x80x9d, Escom, Leiden (1991).
As will be appreciated by those skilled in the art the peptides of the present invention will bind to the TNF receptor. Accordingly, the peptides of the present invention can be used in assays for the presence of TNF receptor in samples. These samples may be either biological fluids or tissue sections. When used in this manner it is preferred that the peptides are labelled with a detectable label.
It will also be appreciated by those skilled in the art that the peptides of the present invention bind to inflammatory cells (e.g. macrophages and neutrophils). The presence of large numbers of these inflammatory cells at a particular site is diagnostic of infectious and non-infectious inflammation.
Accordingly, in a sixth aspect the present invention consists in a method of detecting a site of inflammation in a subject which method comprises administering to the subject a detectably labeled peptide, the peptide comprising a sequence selected from the group consisting of LFKGQGCPSTHVLLTHTISRI (SEQ ID NO. 6), GLYLIYSQVLFKGQG (SEQ ID NO. 10), HVLLTHTISRIAVSYQTKVNLL (SEQ ID NO. 11), PSTHVLLTHTI (SEQ ID NO. 15), PSAHVLLTHTI (SEQ ID NO. 17), and PSTHVLITHTI (SEQ ID NO. 18); and detecting the labeled peptide.
The phrase xe2x80x9csite of inflammationxe2x80x9d is used herein to denote conditions and their locations that occur in a subject due to tissue damage, regardless of the underlying cause or etiology. This tissue damage can result from microbial infection, autoimmune processes or mechanical trauma.
The site of inflammation may be in an internal structure such as in the brain, chest, abdomen, urogenital tract (e.g. prostate), gastrointestinal tract or joints.
There are many different labels and methods of labeling known to those skilled in the art. Examples of the types of labels that can be used in the present invention include radioactive isotopes, paramagnetic isotopes, and compounds that can be imaged by positron emission tomography (PET). Those skilled in the art will know of other suitable labels for the peptides of the present invention, or will be able to ascertain such using routine experimentation. Furthermore, the binding of these labels to the peptide can be done using standard techniques common to those skilled in the art.
In the context of the method of the sixth aspect, the labelled peptide may be administered by any suitable means. In one preferred embodiment, the administering is parenteral. Preferably, the parenteral administration comprises intradermal, subcutaneous, intramuscular, intraperitoneal, or intravenous injection.
In a seventh aspect, the present invention comprises a method of diagnosing a disease state selected from the group consisting of a bacterial infection, a fungal infection, a parasitic infection, a viral infection, Group B streptococcal disease, osteomyelitis, pneumonia, bronchitis, chronic obstructive pulmonary disease, inflammatory bowel disease, atypical appendicitis, Acquired Iunmune Deficiency Syndrome, a granulomatous disease, respiratory distress syndrome, cystic fibrosis and rheumatoid arthritis, which method comprises administering to a subject a detectably labeled peptide, the peptide comprising a sequence selected from the group consisting of LFKGQGCPSTHVLLTHTISRI (SEQ ID NO. 6), GLYLIYSQVLFKGQG (SEQ ID NO. 10), HVLLTHTISRIAVSYQTKVNLL (SEQ ID NO. 11), PSTHVLLTHTI (SEQ ID NO. 15), PSAHVLLTHTI (SEQ ID NO. 17), and PSTHVLITHTI (SEQ ID NO. 18); and detecting the labeled peptide.
In addition to identifying and characterising sites of local inflammation, the method of the present invention can be used to monitor the course of inflammation in a subject. Thus, by measuring an increase or decrease in the size or number of inflammation sites, it is possible to determine whether a particular therapeutic regimen aimed at ameliorating the cause of inflammation, or directed to the inflammatory process itself, is effective.
It will be appreciated that the methods of the sixth and seventh aspects of the present invention are advantageous when compared to prior methods of imaging inflammation. Prior methods have involved the use of labeled white blood cells or labeled antibodies to identify and/or localize inflammation in vivo Use of white blood cells requires handling of blood (which represents hazards to both patient and medical personnel). Labeled antibodies tend to have long half lives which leads to elevated background in imaging diagnostic procedures. It is expected that the use of labeled peptides for detection and/or quantification of inflammatory sites in vivo will avoid or reduce these problems.
In order that the nature of the present invention may be more clearly understood, preferred forms thereof will now be described with reference to the following Examples and Figures.